Inbred sunflower (helianthus annuus) line designated 503a/b

ABSTRACT

The present invention relates to an inbred sunflower line, designated 503A/B. The invention relates to the seeds of inbred sunflower line 503A/B, to the plants of inbred sunflower line 503A/B and to the methods for producing a sunflower plant, either inbred or hybrid, by crossing the inbred line 503A/B with itself or another sunflower line. The invention further relates to methods for producing a sunflower plant containing in its genetic material one or more transgenes and to the transgenic plants produced by that method and to methods for producing other inbred sunflower lines derived from the inbred 503A/B.

CROSS-REFERENCE TO RELATED APPLICATION

This application claims the benefit of U.S. Provisional PatentApplication Ser. No. 61/301,953, filed Feb. 5, 2010, the disclosure ofwhich is hereby incorporated herein in its entirety by this reference.

FIELD OF THE INVENTION

This invention is in the field of sunflower breeding. Specifically thepresent invention relates to new and distinctive sunflower cultivar,designated 503A/B.

BACKGROUND OF THE INVENTION

The cultivated sunflower (Helianthus annus L.) is a major worldwidesource of vegetable oil. In the United States, approximately 4 millionacres are planted in sunflowers annually, primarily in the Dakotas andMinnesota.

Field crops are bred through techniques that take advantage of theplant's method of pollination. A plant is self-pollinated if pollen fromone flower is transferred to the same or another flower of the sameplant. A plant is cross-pollinated if the pollen comes from a flower ona different plant.

Plants that have been self-pollinated and selected for type for manygenerations become homozygous at almost all gene loci and produce auniform population of true breeding progeny. A cross between twodifferent homozygous lines produces a uniform population of hybridplants that may be heterozygous for many gene loci. A cross of twoplants each heterozygous for many gene loci. A cross of two plants eachheterozygous at a number of gene loci will produce a population ofhybrid plants that differ genetically and will not be uniform.

There are numerous steps in the development of any novel, desirableplant germplasm. Plant breeding begins with the analysis and definitionof problems and weaknesses of the current germplasm, the establishmentof program goals, and the definition of specific breeding objectives.The next step is selection of germplasm that possess the traits to meetthe program goals. The goal is to combine in a single variety animproved combination of desirable traits from the parental germplasm.These important traits may include higher seed yield, resistance todiseases and insects, resistance to herbicides, better stems and roots,tolerance to drought and heat, and better agronomic quality.

Choice of breeding or selection methods depends on the mode of plantreproduction, the heritability of the trait(s) being improved, and thetype of cultivar used commercially (e.g., F₁ hybrid cultivar, purelinecultivar, etc.). For highly heritable traits, a choice of superiorindividual plants evaluated at a single location will be effective,whereas for traits with low heritability, selection should be based onmean values obtained from replicated evaluations of families of relatedplants. Popular selection methods commonly include pedigree selection,modified pedigree selection, mass selection, and recurrent selection.

The complexity of inheritance influences choice of the breeding method.Backcross breeding is used to transfer one or a few favorable genes fora highly heritable trait into a desirable cultivar. This approach hasbeen used extensively for breeding disease-resistant cultivars. Variousrecurrent selection techniques are used to improve quantitativelyinherited traits controlled by numerous genes. The use of recurrentselection in self-pollinating crops depends on the ease of pollination,the frequency of successful hybrids from each pollination, and thenumber of hybrid offspring from each successful cross.

Each breeding program should include a periodic, objective evaluation ofthe efficiency of the breeding procedure. Evaluation criteria varydepending on the goal and objectives, but should include gain fromselection per year based on comparisons to an appropriate standard,overall value of the advanced breeding lines, and number of successfulcultivars produced per unit of input (e.g., per year, per dollarexpended, etc.).

Sunflower (Helianthus annus L.), can be bred by both self-pollinationand cross-pollination techniques. The sunflower head (inflorescence)usually is composed of about 1,000 to 2,000 individual disk flowersjoined to a common base (receptacle). The flowers around thecircumference are ligulate ray flowers with neither stamens nor pistil.The remaining flowers are hermaphroditic and protandrous disk flowers.

Natural pollination of sunflower occurs when flowering starts with theappearance of a tube partly exerted from the sympetalous corolla. Thetube is formed by the finve syngenesious anthers, and pollen is releasedon the inner surface of the tube. The style lengthens rapidly and forcesthe stigma through the tube. The two lobes of the stigma open outwardand are receptive to pollen but out of reach of their own polleninitially. Although this largely prevents self-pollination of individualflowers, flowers are exposed to pollen from other flowers on the samehead by insects, wind and gravity.

Promising advanced breeding lines are thoroughly tested and compared toappropriate standards in environments representative of the commercialtarget area(s) for three or more years. The best lines are candidatesfor new commercial cultivars; those still deficient in a few traits maybe used as parents to produce new populations for further selection.

These processes, which lead to the final step of marketing anddistribution, usually take from eight to 12 years from the time thefirst cross is made. Therefore, development of new cultivars is atime-consuming process that requires precise forward planning, efficientuse of resources, and a minimum of changes in direction.

A difficult task is the identification of individuals that aregenetically superior because, for most traits, the true genotypic valueis masked by other confounding plant traits or environmental factors.One method of identifying a superior plant is to observe its performancerelative to other experimental plants and to a widely grown standardcultivar. If a single observation is inconclusive, replicatedobservations provide a better estimate of its genetic worth.

The goal of plant breeding is to develop new, unique and superiorsunflower cultivars and hybrids. The breeder initially selects andcrosses two or more parental lines, followed by repeated selfing andselection, producing many new genetic combinations. The breeder cantheoretically generate billions of different genetic combinations viacrossing, selfing and mutations. The breeder has no direct control atthe cellular level. Therefore, two breeders will never develop the sameline, or even very similar lines, having the same sunflower traits.

Each year, the plant breeder selects the germplasm to advance to thenext generation. This germplasm is grown under unique and differentgeographical, climatic and soil conditions and further selections arethen made, during and at the end of the growing season. The cultivarswhich are developed are unpredictable. This unpredictability is due tothe breeder's selection, which occurs in unique environments, with nocontrol at the DNA level (using conventional breeding procedures), andwith millions of different possible genetic combinations beinggenerated. A breeder of ordinary skill in the art cannot predict thefinal resulting lines he develops, except possibly in a very gross andgeneral fashion. The same breeder cannot produce the same cultivar twiceby using the exact same original parents and the same selectiontechniques. This unpredictability results in the expenditure of largeamounts of research monies to develop superior new sunflower cultivars.

The development of new sunflower cultivars requires the development andselection of sunflower varieties, the crossing of these varieties, andselection of superior hybrid crosses. The hybrid seed is produced bymanual crosses between selected male-fertile parents or by using malesterility systems. These hybrids are selected for certain single genetraits such as pod color, flower color, pubescence color, or herbicideresistance which indicate that the seed is truly a hybrid. Additionaldata on parental lines, as well as the phenotype of the hybrid,influence the breeder's decision whether to continue with the specifichybrid cross.

Pedigree breeding and recurrent selection breeding methods are used todevelop cultivars from breeding populations. Breeding programs combinedesirable traits from two or more cultivars or various broad-basedsources into breeding pools from which cultivars are developed byselfing and selection desired phenotypes. The new cultivars areevaluated to determine which have commercial potential.

Pedigree breeding is used commonly for the improvement ofself-pollinating crops. Two parents which possess favorable,complementary traits are crossed to produce an F₁. An F₂ population isproduced by selfing one or several F₁'s. Selection of the bestindividuals may begin in the F₂ population; then, beginning in the F₃,the best individuals in the best families are selected. Replicatedtesting of families can begin in the F₄ generation to improve theeffectiveness of selection for traits with low heritability. At anadvanced stage of inbreeding (i.e., F₆ and F₇), the best lines ormixtures of phenotypically similar lines are tested for potentialrelease as new cultivars.

Mass and recurrent selection can be used to improve populations ofeither self- or cross-pollinating crops. A genetically variablepopulation of heterozygous individuals is either identified or createdby intercrossing several different parents. The best plants are selectedbased on individual superiority, outstanding progeny, or excellentcombining ability. The selected plants are intercrossed to produce a newpopulation in which further cycles of selection are continued.

Backcross breeding has been used to transfer genes for simply inheritedhighly heritable trait into a desirable homozygous cultivar or inbredline which is the recurrent parent. The source of the trait to betransferred is called the donor parent. The resulting plant is expectedto have the attributes of the recurrent parent (e.g., cultivar) and thedesirable trait transferred from the donor parent. After the initialcross, individuals possessing the phenotype of the donor parent areselected and repeatedly crossed (backcrossed) to the recurrent parent.The resulting plant is expected to have the attributes of the recurrentparent (e.g., cultivar) and the desirable trait transferred from thedonor parent.

The single-seed descent procedure in the strict sense refers to plantinga segregating population, harvesting a sample of one seed per plant, andusing the one-seed sample to plant the next generation. When thepopulation has been advanced from the F₂ to the desired level ofinbreeding, the plants from which lines are derived will each trace todifferent F₂ individuals. The number of plants in a population declineseach generation due to failure of some seeds to germinate or some plantsto produce at least one seed. As a result, not all of the F₂ plantsoriginally sampled in the population will be represented by a progenywhen generation advance is completed.

In a multiple-seed procedure, sunflower breeders commonly harvest seedsfrom each plant in a population and thresh them together to form a bulk.Part of the bulk is used to plant the next generation and part is put inreserve. The procedure has been referred to as modified single-seeddescent.

The multiple-seed procedure has been used to save labor at harvest. Itis considerably faster to remove seeds with a machine than to remove oneseed from each by hand for the single-seed procedure. The multiple-seedprocedure also makes it possible to plant the same number of seeds of apopulation each generation of inbreeding. Enough seeds are harvested tomake up for those plants that did not germinate or produce seed.

Descriptions of other breeding methods that are commonly used fordifferent traits and crops can be found in one of several referencebooks (e.g., Allard, 1960; Simmonds, 1979; Sneep et al., 1979; Fehr,1987), the contents of which are incorporated herein by this reference.

Proper testing should detect any major faults and establish the level ofsuperiority or improvement over current cultivars. In addition toshowing superior performance, there must be a demand for a new cultivarthat is compatible with industry standards or which creates a newmarket. The introduction of a new cultivar can incur additional costs tothe seed producer, the grower, processor and consumer due to specialadvertising and marketing, altered seed and commercial productionpractices, and new product utilization. The testing preceding release ofa new cultivar should take into consideration research and developmentcosts as well as technical superiority of the final cultivar. Forseed-propagated cultivars, it must be feasible to produce seed easilyand economically.

Once the inbred plants that give the best hybrid performance have beenidentified, the hybrid seed can be reproduced indefinitely as long asthe homogeneity of the inbred parent is maintained. A single-crosshybrid is produced when two inbred lines are crossed to produce the F1progeny. A double-cross hybrid is produced from four inbred linescrossed in pairs (A×B and C×D) and then the two F1 hybrids are crossedagain (A×B)×(C×D). Much of the hybrid vigor exhibited by F1 hybrids islost in the next generation (F2). Consequently, seed from hybridvarieties is not used for planting stock.

The very rapid expansion over the last decade of acreage planted withsunflower in the United States is due in part to several importantdevelopments in the field of sunflower breeding and varietalimprovement. On significant development was the discovery of cytoplasmicmale sterility and genes for fertility restoration, a discovery thatallowed the production of hybrid sunflowers. The hybrids thus producedwere introduced during the early 1970's. A description of cytoplasmicmale sterility (CMS) and genetic fertility restoration in sunflowers ispresented by Fick, “Breeding and Genetics,” in Sunflower Science andTechnology 279-338 (J. F. Carter ed. 1978), the contents of which areincorporated herein by this reference.

A reliable method of controlling male fertility in plants offers theopportunity for improved plant breeding. This is especially true fordevelopment of sunflower hybrids, which relies upon some sort of malesterility system. Two types of male sterility, genetic and cytoplasmic,have been found in sunflower. The use of male sterility in plantbreeding has been described in U.S. Pat. No. 6,956,156, the contents ofwhich are incorporated herein by this reference.

Sunflower, Helianthus annus L., is an important and valuable field crop.Thus, a continuing goal of plant breeders is to develop stable, highyielding sunflower cultivars that are agronomically sound. A currentgoal is to maximize the amount of grain produced on the land used and tosupply food for both animals and humans. To accomplish this goal, thesunflower breeder must select and develop sunflower plants that havetraits that result in superior cultivars.

Weed species have long been a problem in cultivated fields. Althoughonce a labor intensive operation, weed control has been made easier bythe availability of efficient weed killing chemical herbicides. Thewide-spread use of herbicides, along with improved crop varieties andfertilizers, has made a significant contribution to the “greenrevolution” in agriculture. Not all herbicides are capable ofselectively targeting the undesirable plants over crop plants, as wellas being non-toxic to animals. Often it is necessary to settle forcompounds which are simply less toxic to crop plants than to weeds.Particularly useful herbicides are those that have a broad spectrum ofherbicidal activity. Unfortunately, broad spectrum herbicides typicallyhave deleterious effect on crop plants exposed to the herbicide. As suchthe development of herbicide resistant crop plants has become a majorfocus of agricultural research.

On particular broad spectrum herbicide that has been investigated isimidazolinone. The imidazolinone herbicides include: imazapyr, imazapic,imazethapyr, imazamox, imazamethabenz, and imazaquin. These herbicidescontrol weeds by disrupting the activity of the enzyme acetohydroxyacidsynthase (AHAS) also called acetolactate synthase (ALS). AHAS is acritical enzyme for the biosynthesis of branched chain amino acids inplants, as is described in Tan et al. (2005), Imidazolinone-tolerantcrops: history, current status, and future, Pest Management Science,vol. 61: pp 246-257, the contents of which are incorporated herein bythis reference. There are several variant AHAS genes which haveconferred imidazolinone tolerance and have been used to create variousimidazolinone-tolerant crops, as has been described in U.S. Pat. Nos.5,767,361, and 4,761,373, which are both incorporated herein by thisreference. Mutations in the AHAS coding regions alter the enzymestructure and prevent inhibition of the enzyme by the herbicide.Tolerance to broad spectrum herbicides provides an economically viablemethod to control a wide range of weeds in domesticated crops.

Disease in plants is caused by biotic and abiotic causes. Biotic causesof disease include fungi, viruses, bacteria, and nematodes. Of these,fungi are the most frequent causative agent of disease in plants. Thefungus causing downy mildew of cultivated sunflowers, also known asPlasmopara halstedii is a major pathogen affecting domesticatedsunflower crops. The various hosts for downy mildew have been describedin E. E. Leppik (1966) Origin and specialization of Plasmopara halstediicomplex on Compositae. FAO Plant Protection Bulletin 14, 72-76, and N.S. Novotel'nova (1966) [Downy mildew of sunflower], 150 pp. Nauka,Moscow, Russia, the contents of which are incorporated herein by thisreference. Downy mildew is a soil-borne pathogen, inoculating youngsunflower seedlings primarily with its oospores. There is also thechance for wind-borne infection which is spread via sporangia, or can beseed-borne from infected plants, but usually only leads to limitedlocalized infection.

The foregoing examples of the related art and limitations relatedtherewith are intended to be illustrative and not exclusive. Otherlimitations of the related art will become apparent to those of skill inthe art upon a reading of the specification.

BRIEF SUMMARY OF THE INVENTION

The following embodiments and aspects thereof are described inconjunction with systems, tools and methods which are meant to beexemplary and illustrative, not limiting in scope. In variousembodiments, one or more of the above-described problems have beenreduced or eliminated, while other embodiments are directed to otherimprovements.

According to the invention, there is provided a novel inbred sunflowerline, designated 503A/B. Particular embodiments of the invention relateto the seeds of inbred sunflower line 503A/B, to the plants of inbredsunflower line 503A/B and to methods for producing a sunflower plantproduced by crossing the inbred line 503A/B with itself or anothersunflower line, and to methods for producing a sunflower plantcontaining in its genetic material one or more transgenes and to thetransgenic sunflower plants produced by that method. Other embodimentsrelate to methods for producing other inbred sunflower lines derivedfrom inbred sunflower 503A/B and to the inbred sunflower lines derivedby the use of those methods. Additional embodiments of the inventionfurther relate to hybrid sunflower seeds and plants produced by crossingthe inbred line 503A/B with another sunflower line.

The inbred sunflower plant of the invention may further comprise, orhave, a cytoplasmic factor that is capable of conferring male sterility.Parts of the sunflower plant of the present invention are also provided,such as, e.g., pollen obtained from an inbred plant and an ovule of theinbred plant.

In another aspect, the present invention provides regenerable cells foruse in tissue culture or inbred sunflower plant 503A/B. The tissueculture can be capable of regenerating plants having the physiologicaland morphological characteristics of the foregoing inbred sunflowerplant. The regenerable cells in such tissue cultures can be embryos,pollen, ovules, leaves, stems, cortex, pith, involucral bracts, rayflowers, disk flowers, pappi, achenes, nectarines, interfloral bracts,receptacle, trichomes stigma, anther, style, filament, calyx, pericarp,seed coat, endosperm, embryo, roots, root tips, and seeds. Additionally,the present invention provides sunflower plants regenerated from thetissue cultures of the invention.

DETAILED DESCRIPTION OF THE INVENTION

In the description and tables which follow, a number of terms are used.In order to provide a clear and consistent understanding of thespecification and claims, including the scope to be given such terms,the following definitions are provided:

Allele. Allele is any of one or more alternative forms of a gene, all ofwhich alleles relate to one trait or characteristic. In a diploid cellor organism, the two alleles of a given gene occupy corresponding locion a pair of homologous chromosomes.

Backcrossing. Backcrossing is a process in which a breeder repeatedlycrosses hybrid progeny back to one of the parents, for example, a firstgeneration hybrid F₁ with one of the parental genotypes of the F₁hybrid.

Cytoplasmic Male Sterile (CMS) Plant or Inbred Line. A sunflower linethat produces no viable pollen is called male sterile. Male sterility isinherited maternally, i.e., the male sterile plant is used as the femaleparent in a cross with pollen from another sunflower. CMS lines areproduced by crossing a maintainer line with a sunflower plant with thecytoplasmic male sterility trait and then backcrossing to the maintainerline until a male sterile line that is homologous to the maintainer linein all other respects is developed. CMS lines are also referred to asfemale lines.

Elite sunflower. A sunflower cultivar which has been stabilized forcertain commercially important agronomic traits comprising a stabilizedyield of about 100% or greater relative to the yield of check varietiesin the same growing location growing at the same time and under the sameconditions. In one embodiment, “elite sunflower” means a sunflowercultivar stabilized for certain commercially important agronomic traitscomprising a stabilized yield of 110% or greater relative to the yieldof check varieties in the same growing location growing at the same timeand under the same conditions. In another embodiment, “elite sunflower”means a sunflower cultivar stabilized for certain commercially importantagronomic traits comprising a stabilized yield of 115% or greaterrelative to the yield of check varieties in the same growing locationgrowing at the same time and under the same conditions.

Elite sunflower cultivar. A sunflower cultivar, per se, which has beensold commercially.

Elite sunflower parent cultivar. A sunflower cultivar which is theparent cultivar of a canola hybrid that has been commercially sold.

Embryo. The embryo is the small plant contained within a mature seed.

FAME analysis. Fatty Acid Methyl Ester analysis is a method that allowsfor accurate quantification of the fatty acids that make up complexlipid classes.

Glucosinolates. These are measured in micromoles (μm) of total alipathicglucosinolates per gram of air-dried oil-free meal. The level ofglucosinolates is somewhat influenced by the sulfur fertility of thesoil, but is also controlled by the genetic makeup of each variety andthus can be useful in characterizing varieties.

Imidazolinone resistance (Imi). Resistance and/or tolerance is conferredby one or more genes which alter acetolactate synthase (ALS), also knownas acetohydroxy acid synthase (AHAS) allowing the enzyme to resist theaction of imidazolinone.

Leaf blade color. The color of the leaf blades is variety specific andcan range from light to medium dark green to blue green.

Leaf development of lobes. The leaves on the upper portion of the stemcan show varying degrees of development of lobes which are disconnectedfrom one another along the petiole of the leaf. The degree of lobing isvariety specific and can range from absent (no lobes)/weak through verystrong (abundant lobes).

Leaf indentation of margin. The leaves on the upper portion of the stemcan also show varying degrees of serration along the leaf margins. Thedegree of serration or indentation of the leaf margins can vary fromabsent (smooth margin)/weak to strong (heavy saw-tooth like margin).

Leaf surface. The leaf surface can also be used to distinguish betweenvarieties. The surface can be smooth or rugose (lumpy) with varyingdegrees between the two extremes.

Maturity or Date to Maturity. The maturity of a variety is measured asthe number of days between planting and physiological maturity. This isuseful trait in distinguishing varieties relative to one another.

Mutagenesis. Mutagenesis refers to mutagenesis of a plant or plant partwith a mutagen (e.g., a chemical or physical agent that increases thefrequency of mutations in a target plant or plant part). By way ofnon-limiting example, the double chemical mutagenesis technique ofKonzak, as described in U.S. Pat. No. 6,696,294 the contents of whichare incorporated herein by this reference, can be used to induce mutantalleles in endogenous plant genes.

Percent linolenic acid. Percent oil of the seed that is linolenic acid.

Oil content. This is measured as percent of the whole dried seed and ischaracteristic of different varieties. It can be determined usingvarious analytical techniques such as NMR, NIR, and Soxhlet extraction.

Percent oleic acid (OLE). Percent oil of the seed that is oleic acid.

Percentage of total fatty acids. This is determined by extracting asample of oil from seed, producing the methyl esters of fatty acidspresent in that oil sample and analyzing the proportions of the variousfatty acids in the sample using gas chromatography. The fatty acidcomposition can also be a distinguishing characteristic of a variety.

Petal color. The petal color on the first day a flower opens can be adistinguishing characteristic for a variety. It can be white, varyingshades of yellow or orange.

Plant height. This is the height of the plant at the end of flowering ifthe floral branches are extended upright (i.e., not lodged). This variesfrom variety to variety and although it can be influenced byenvironment, relative comparisons between varieties grown side by sideare useful for variety identification.

Protein content. This is measured as percent of whole dried seed and ischaracteristic of different varieties. This can be determined usingvarious analytical techniques such as NIR and Kjeldahl.

Quantitative Trait Loci (QTL). Quantitative trait loci (QTL) refer togenetic loci that control to some degree numerically representabletraits that are usually continuously distributed.

Regeneration. Regeneration refers to the development of a plant fromtissue culture.

Restorer Line. A line possessing the gene or genes to restore malefertility or viable pollen to a sunflower hybrid or inbred line andprogeny having a maternal cytoplasm that conditions male sterility.

Single Gene Converted (Conversion). Single gene converted (conversion)plant refers to plants which are developed by a plant breeding techniquecalled backcrossing, or via genetic engineering, wherein essentially allof the desired morphological and physiological characteristics of avariety are recovered in addition to the single gene transferred intothe variety via the backcrossing technique or via genetic engineering.

Stabilized. Reproducibly passed from one generation to the nextgeneration of inbred plants of same variety.

Total Saturated (TOTSAT). Total percent oil of the seed of the saturatedfats in the oil including C12:0, C14:0, C16:0, C18:0, C20:0, C22:0 andC24.0.

Mean Yield. Mean yield of all canola entries grown at a given location.

Yield. Greater than 10% above the mean yield across 10 or morelocations.

Check Average. Average for one or more checks in a given location.

Some of the criteria used to select in various generations include: seedyield, lodging resistance, emergence, disease tolerance, maturity, lateseason plant intactness, plant height and shattering resistance.

The cultivar has shown uniformity and stability, as described in thefollowing variety description information. It has been self-pollinated asufficient number of generations with careful attention to uniformity ofplant type. The cultivar has been increased with continued observationfor uniformity.

Seed from sunflower line 503A/B has been deposited with the AmericanType Culture Collection (ATCC), 12301 Parklaw Drive, Rockville, Md., USA20852 and bearing ATCC Accession Number PTA-10098. Inbred sunflower line503A/B is an oil type sunflower male line with superior characteristics,and provides an excellent parental line in crosses for producing firstgeneration (F₁) hybrid sunflower.

Inbred sunflower line 503A/B has the following morphologic and othercharacteristics, as described in Table 1.

TABLE 1 Characteristic Value CLASS (1 = Oil Type, or 2 = Confectionery,non-oil type) 1 NO. OF DAYS TO FLOWERING 65 NO. OF DAYS TO MATURITY 95PLANT HEIGHT AT MATURITY (cm) 76 NUMBER OF LEAVES AT FLOWERING 27 STEMBRANCHING 1 = No Branching, 2 = Basal Branching, 3 = Top Branching 1(with central head), and 4 = Fully Branched (without central head).INTERNODE LENGTH AT FLOWERING (cm) 2.3 STEM COLOR OF GROWING POINT (1 =Green or 2 = Yellow) 1 DEPTH OF LEAF MARGIN INDENTATIONS (1 = shallow, 2= intermediate, or 2 3 = deep) LEAF APEX (1 = acuminate or 2 = other) 1LEAF ATTITUDE (1 = erect, 2 = ascending, 3 = horizontal or 4 =descending) 2 LEAF BASE 1 = auriculate, 2 = truncate, 3 = acute, 4 =rounded, or 5 = cordate 1 LEAF BLADE LENGTH (cm) 20. LEAF BLADE WIDTH(cm) 21 LEAF COLOR (1 = light green, 2 = green, 3 = dark green, or 4 =brown) 2 LEAF MARGIN (1 = entire, 2 = crenate, or 3 = serrate) 2 LEAFMARGIN COLOR (1 = green or 2 = yellow) 1 LEAF SHAPE (1 = cordate, 2 =lanceolate, 3 = triangular, or 4 = round) 1 LEAF SURFACE (1 = smooth, 2= crinkled (ridged) or 3 = other) 2 LEAF WIDTH:LENGTH RATIO (1 =narrower than long, 2 = equal or 3 = wider than long) 3 RAY FLOWERS(presence or absence) Present RAY FLOWER COLOR (1 = yellow, 2 = sulfuryellow, 3 = orange yellow or 4 = other) 1 DISK FLOWER COLOR (1 = yellow,2 = red, or 3 = purple) 1 ANTHOCYANIN IN STIGMAS (presence or absence)PAPPI COLOR (1 = green or 2 = rust (red)) POLLEN COLOR (1 = white(colorless) or 2 = yellow) 2 RAY LENGTH (mm) RAY WIDTH (mm) HEADATTITUDE (1 = vertical (erect), 2 = ascending, 3 = horizontal, or 1 4 =descending) HEAD DIAMETER (cm) 20. HEAD RECEPTACLE SHAPE (1 = flat, 2 =convex, or 3 = concave) NO. OF SEEDS PER HEAD OUTER PERICARP (1 = clear,2 = striped black, 3 = nearly solid black) 3 MIDDLE PERICARP (1 = whiteor 2 = solid purple) 1 INNER PERICARP (1 = no color or 2 = brownishblack) 1 SEED LENGTH (mm) 10 SEED MOTTLING (1 = absent or 2 = present) 1SEED SHAPE (1 = ovate, 2 = obovate (shield), 3 = narrowly obovate, 4 =oblong or 2 5 = elliptic) SEED SHAPE (CROSS SECTION) (1 = not curved or2 = curved) 2 SEED SIZE (% Held on 7.9 mm (20/64) Round-hole Screen) 0SEED STRIPES (1 = absent, 2 = even black and white stripes, 3 = broadblack and 4 narrow white stripes, 4 = black with narrow dark-greystriping or 5 = other) SEED WEIGHT (gm) 49.1/1000 sds BROOM RAPE (1 =susceptible, or 2 = resistant) 1 CHARCOAL ROT (1 = susceptible, or 2 =resistant) Not tested DOWNY MILDEW (1 = susceptible, or 2 = resistant) 1EUROPEAN SUNFLOWER MOTH (1 = susceptible, or 2 = resistant) Not testedGRAY-MOLD BLIGHT (1 = susceptible, or 2 = resistant) Not tested N.AMERICAN HEAD MOTH (1 = susceptible, or 2 = resistant) Not tested RUST(1 = susceptible, or 2 = resistant) Not tested SCLEROTINIA WILT (1 =susceptible, or 2 = resistant) Not tested VERTICILLIUM WILT (1 =susceptible, or 2 = resistant) Not tested WHITE BLISTER RUST (1 =susceptible, or 2 = resistant) Not tested HULL (%) Not tested LINOLEICACID (%) OIL (%) 41.1 OLEIC ACID (%) 86.1 PALMITIC ACID (%) 4.8 STEARICACID (%) 5.9 TOTAL SATURATES (%) The yield of inbred line s671 (503A X319R) is 2399 kg/ha.

This invention is also direct to methods for producing a sunflower plantby crossing a first parent sunflower plant with a second parentsunflower plant, wherein the first or seconds sunflower plant is theinbred sunflower plant from the line 503A/B. Further, both first andsecond parent sunflower plants may be from the inbred line 503A/B.Therefore, any methods using the inbred sunflower line 503A/B are partof this invention: selfing, backcrosses, hybrid breeding, and crosses topopulations. Any plants produced using inbred sunflower line 503A/B as aparent are within the scope of this invention. Advantageously, theinbred sunflower line is used in crosses with other sunflower varietiesto produce first generation (F₁) sunflower hybrid seed and plants withsuperior characteristics.

503A/B has seeds of approximately 40-45% oil content (at 10% moisturebasis) with 86-90% oleic acid (18:1), and a total saturated fatty acidcontent of approximately 7.0-8.5%.

Some of the criteria used to select plants in various generationsinclude: seed yield, maturity, plant height, uniformity of plant type,disease and insect resistance, and large seed size. During thedevelopment of the line, crosses were made to inbred testers for thepurpose of estimating the line's general and specific combining ability.The inbred was evaluated further as a line and in numerous combinationsacross the Sunflower Belt. The inbred has proven to have a very goodcombining ability in hybrid combinations.

503A/B is an inbred sunflower line that was developed from the cross of500B(1)/68204B through traditional plant breeding backcross methodology.The population cross was designated 500B[1]/68204B and the root linederivation is 500B[1]/68204B-5-5-6-1-2-1-BK. Inbred 503A/B appearsstable and uniform after one generation of backcrossing and hasexhibited commercial value as a parent of short stature sunflowerhybrids in field evaluations. This inbred has potential commercial valueas a parent of sunflower hybrids in field evaluations. The most advancedhybrids with this line as a parent are s671 (503A/319R), s674(503A/318R), and s878 (503A/222R).

503A/B is a high oleic oil seed dwarf line. It contributes larger seedsize and increased seed length relative to previously developedconfection restorers. In addition, seed color and standability ofhybrids using 503A/B is improved. It is a group 4 maturing inbred ofmedium tall plant height with good pollen production for use in hybridproduction fields. 503A/B has shown good general combining ability andyield stability in various hybrid combinations under a wide spectrum ofenvironments. It has been observed to be broadly adapted, showing goodperformance in Argentina, and all sunflower production areas of theUnited States.

The inbred has shown uniformity and stability within the limits ofenvironmental influence for all of the traits. The inbred has beenself-pollinated and a sufficient number of generations, with carefulattention to uniformity of plant type to ensure homozygosity andphenotypic stability necessary to use in commercial production. The linehas been increased both by hand and sibbed in isolated fields withcontinued observations for uniformity. No variant traits have beenobserved or are expected in 503A/B.

Expression Vectors for Sunflower Transformation: Marker Genes

Expression vectors include at least one genetic marker, operably linkedto a regulatory element (a promoter, for example) that allowstransformed cells containing the marker to be either recovered bynegative selection, i.e., inhibiting growth of cells that do not containthe selectable marker gene, or by positive selection, i.e., screeningfor the product encoded by the genetic marker. Many commonly usedselectable marker genes for plant transformation are well known in thetransformation arts, and include, for example, genes that code forenzymes that metabolically detoxify a selective chemical agent which maybe an antibiotic or an herbicide, or genes that encode an altered targetwhich is insensitive to the inhibitor. A few positive selection methodsare also known in the art.

One commonly used selectable marker gene for plant transformation is theneomycin phosphotransferase II (nptII) gene under the control of plantregulatory signals which confers resistance to kanamycin. Fraley et al.,Proc. Natl. Acad. Sci. U.S.A., 80:4803 (1983). Another commonly usedselectable marker gene is the hygromycin phosphotransferase gene whichconfers resistance to the antibiotic hygromycin. Vanden Elzen et al.,Plant Mol. Biol., 5:299 (1985).

Additional selectable marker genes of bacterial origin that conferresistance to antibiotics include gentamycin acetyl transferase,streptomycin phosphotransferase, aminoglycoside-3′-adenyl transferaseand the bleomycin resistance determinant Hayford et al., Plant Physiol.86:1216 (1988); Jones et al., Mol. Gen. Genet., 210:86 (1987); Svab etal., Plant Mol. Biol. 14:197 (1990); Hille et al., Plant Mol. Biol.7:171 (1986). Other selectable marker genes confer resistance toherbicides such as glyphosate, glufosinate, or bromoxynil. Comai et al.,Nature 317:741-744 (1985); Gordon-Kamm et al., Plant Cell 2:603-618(1990); and Stalker et al., Science 242:419-423 (1988).

Other selectable marker genes for plant transformation are not ofbacterial origin. These genes include, for example, mouse dihydrofolatereductase, plant 5-enolpyruvylshikimate-3-phosphate synthase and plantacetolactate synthase. Eichholtz et al., Somatic Cell Mol. Genet. 13:67(1987); Shah et al., Science 233:478 (1986); Charest et al., Plant CellRep. 8:643 (1990).

Another class of marker genes for plant transformation requiresscreening of presumptively transformed plant cells rather than directgenetic selection of transformed cells for resistance to a toxicsubstance, such as an antibiotic. These genes are particularly useful toquantify or visualize the spatial pattern of expression of a gene inspecific tissues and are frequently referred to as reporter genesbecause they can be fused to a gene or gene regulatory sequence for theinvestigation of gene expression. Commonly used genes for screeningpresumptively transformed cells include β-glucuronidase (GUS),β-galactosidase, luciferase and chloramphenicol acetyltransferase. R. A.Jefferson, Plant Mol. Biol. Rep. 5:387 (1987); Teeri et al., EMBO J.8:343 (1989); Koncz et al., Proc. Natl. Acad. Sci. U.S.A. 84:131 (1987);DeBlock et al., EMBO J. 3:1681 (1984).

Recently, in vivo methods for visualizing GUS activity that do notrequire destruction of plant tissue have been made available. MolecularProbes publication 2908, Imagene, T. M. Green, p. 1-4 (1993) and Nalewayet al., J. Cell Biol. 115:151a (1991). However, these in vivo methodsfor visualizing GUS activity have not proven useful for recovery oftransformed cells because of low sensitivity, high fluorescentbackgrounds and limitations associated with the use of luciferase genesas selectable markers.

More recently, a gene encoding Green Fluorescent Protein (GFP) has beenutilized as a marker for gene expression in prokaryotic and eukaryoticcells. Chalfie et al., Science 263:802 (1994). GFP and mutants of GFPmay be used as screenable markers.

Expression Vectors for Sunflower Transformation: Promoters

Genes included in expression vectors must be driven by a nucleotidesequence comprising a regulatory element, for example, a promoter.Several types of promoters are now well known in the transformationarts, as are other regulatory elements that can be used alone or incombination with promoters.

As used herein, “promoter” includes reference to a region of DNAupstream from the start of transcription and involved in recognition andbinding of RNA polymerase and other proteins to initiate transcription.A “plant promoter” is a promoter capable of initiating transcription inplant cells. Examples of promoters under developmental control includepromoters that preferentially initiate transcription in certain tissues,such as leaves, roots, seeds, fibers, xylem vessels, tracheids, orsclerenchyma. Such promoters are referred to as “tissue-preferred.”Promoters which initiate transcription only in certain tissues arereferred to as “tissue-specific.” A “cell type” specific promoterprimarily drives expression in certain cell types in one or more organs,for example, vascular cells in roots or leaves. An “inducible” promoteris a promoter which is under environmental control. Examples ofenvironmental conditions that may effect transcription by induciblepromoters include anaerobic conditions or the presence of light.Tissue-specific, tissue-preferred, cell type specific, and induciblepromoters constitute the class of “non-constitutive” promoters. A“constitutive” promoter is a promoter which is active under mostenvironmental conditions.

A. Inducible Promoters

An inducible promoter is operably linked to a gene for expression insunflower. Optionally, the inducible promoter is operably linked to anucleotide sequence encoding a signal sequence which is operably linkedto a gene for expression in sunflower. With an inducible promoter, therate of transcription increases in response to an inducing agent.

Any inducible promoter can be used in the instant invention. See Ward etal., Plant Mol. Biol. 22:361-366 (1993). Exemplary inducible promotersinclude, but are not limited to, that from the ACEI system whichresponds to copper (Mett et al., PNAS 90:4567-4571 (1993)); In2 genefrom maize which responds to benzenesulfonamide herbicide safeners(Hershey et al., Mol. Gen. Genetics 227:229-237 (1991); and Gatz et al.,Mol. Gen. Genetics 243:32-38 (1994)) or Tet repressor from Tn10 (Gatz etal., Mol. Gen. Genetics 227:229-237 (1991)). A particularly preferredinducible promoter is a promoter that responds to an inducing agent towhich plants do not normally respond. An exemplary inducible promoter isthe inducible promoter from a steroid hormone gene, the transcriptionalactivity of which is induced by a glucocorticosteroid hormone. Schena etal., Proc. Natl. Acad. Sci. U.S.A. 88:0421 (1991).

B. Constitutive Promoters

A constitutive promoter is operably linked to a gene for expression insunflower or the constitutive promoter is operably linked to anucleotide sequence encoding a signal sequence which is operably linkedto a gene for expression in sunflower.

Many different constitutive promoters can be utilized in the instantinvention. Exemplary constitutive promoters include, but are not limitedto, the promoters from plant viruses such as the 35S promoter from CaMV(Odell et al., Nature 313:810-812 (1985)) and the promoters from suchgenes as rice actin (McElroy et al., Plant Cell 2:163-171 (1990));ubiquitin (Christensen et al., Plant Mol. Biol. 12:619-632 (1989); andChristensen et al., Plant Mol. Biol. 18:675-689 (1992)); pEMU (Last etal., Theor. Appl. Genet. 81:581-588 (1991)); MAS (Velten et al., EMBO J.3:2723-2730 (1984)) and maize H3 histone (Lepetit et al., Mol. Gen.Genetics 231:276-285 (1992); and Atanassova et al., Plant Journal 2(3):291-300 (1992)).

C. Tissue-specific or Tissue-Preferred Promoters

A tissue-specific promoter is operably linked to a gene for expressionin sunflower. Optionally, the tissue-specific promoter is operablylinked to a nucleotide sequence encoding a signal sequence which isoperably linked to a gene for expression in sunflower. Plantstransformed with a gene of interest operably linked to a tissue-specificpromoter produce the protein product of the transgene exclusively, orpreferentially, in a specific tissue.

Any tissue-specific or tissue-preferred promoter can be utilized in theinstant invention. Exemplary tissue-specific or tissue-preferredpromoters include, but are not limited to, a root-preferredpromoter—such as that from the phaseolin gene (Murai et al., Science23:476-482 (1983); and Sengupta-Gopalan et al., Proc. Natl. Acad. Sci.U.S.A. 82:3320-3324 (1985)); a leaf-specific and light-induced promotersuch as that from cab or rubisco (Simpson et al., EMBO J.4(11):2723-2729 (1985); and Timko et al., Nature 318:579-582 (1985)); ananther-specific promoter such as that from LAT52 (Twell et al., Mol.Gen. Genetics 217:240-245 (1989)); a pollen-specific promoter such asthat from Zm13 (Guerrero et al., Mol. Gen. Genetics 244:161-168 (1993))or a microspore-preferred promoter such as that from apg (Twell et al.,Sex. Plant Reprod. 6:217-224 (1993)).

Transport of protein produced by transgenes to a subcellular compartmentsuch as the chloroplast, vacuole, peroxisome, glyoxysome, cell wall ormitochondrion or for secretion into the apoplast, is accomplished bymeans of operably linking the nucleotide sequence encoding a signalsequence to the 5′ and/or 3′ region of a gene encoding the protein ofinterest. Targeting sequences at the 5′ and/or 3′ end of the structuralgene may determine, during protein synthesis and processing, where theencoded protein is ultimately compartmentalized.

The presence of a signal sequence directs a polypeptide to either anintracellular organelle or subcellular compartment, or for secretion tothe apoplast. Many signal sequences are known in the art. See, forexample Becker et al., Plant Mol. Biol. 20:49 (1992); P. S. Close,Master's Thesis, Iowa State University (1993); C. Knox et al.,“Structure and Organization of Two Divergent Alpha-Amylase Genes fromBarley,” Plant Mol. Biol. 9:3-17 (1987); Lerner et al., Plant Physiol.91:124-129 (1989); Fontes et al., Plant Cell 3:483-496 (1991); Matsuokaet al., Proc. Natl. Acad. Sci. 88:834 (1991); Gould et al., J. Cell.Biol. 108:1657 (1989); Creissen et al., Plant J. 2:129 (1991); Kalderon,et al., A short amino acid sequence able to specify nuclear location,Cell 39:499-509 (1984); Steifel, et al., Expression of a maize cell wallhydroxyproline-rich glycoprotein gene in early leaf and root vasculardifferentiation, Plant Cell 2:785-793 (1990).

Foreign Protein Genes and Agronomic Genes

With transgenic plants according to the present invention, a foreignprotein can be produced in commercial quantities. Thus, techniques forthe selection and propagation of transformed plants, which are wellunderstood in the art, yield a plurality of transgenic plants which areharvested in a conventional manner, and a foreign protein then can beextracted from a tissue of interest or from total biomass. Proteinextraction from plant biomass can be accomplished by known methods whichare discussed, for example, by Heney and Orr, Anal. Biochem. 114:92-6(1981).

According to a particular embodiment, the transgenic plant provided forcommercial production of foreign protein is a sunflower plant. Inanother preferred embodiment, the biomass of interest is seed. For therelatively small number of transgenic plants that show higher levels ofexpression, a genetic map can be generated, primarily via conventionalRFLP, PCR and SSR analysis, which identifies the approximate chromosomallocation of the integrated DNA molecule. For exemplary methodologies inthis regard, see Glick and Thompson, Methods in Plant Molecular Biologyand Biotechnology, CRC Press, Boca Raton 269:284 (1993). Map informationconcerning chromosomal location is useful for proprietary protection ofa subject transgenic plant. If unauthorized propagation is undertakenand crosses made with other germplasm, the map of the integration regioncan be compared to similar maps for suspect plants, to determine if thelatter have a common parentage with the subject plant. Map comparisonswould involve hybridizations, RFLP, PCR, SSR and sequencing, all ofwhich are conventional techniques.

Likewise, by means of the present invention, agronomic genes can beexpressed in transformed plants. More particularly, plants can begenetically engineered to express various phenotypes of agronomicinterest. Exemplary genes implicated in this regard include, but are notlimited to, those categorized below:

1. Genes That Confer Resistance to Pests or Disease and That Encode:

A. Plant disease resistance genes. Plant defenses are often activated byspecific interaction between the product of a disease resistance gene(R) in the plant and the product of a corresponding avirulence (Avr)gene in the pathogen. A plant variety can be transformed with clonedresistance genes to engineer plants that are resistant to specificpathogen strains. See, for example, Jones et al., Science 266:789 (1994)(cloning of the tomato Cf-9 gene for resistance to Cladosporium fulvum);Martin et al., Science 262:1432 (1993) (tomato Pto gene for resistanceto Pseudomonas syringae pv. tomato encodes a protein kinase); Mindrinoset al., Cell 78:1089 (1994) (Arabidopsis RSP2 gene for resistance toPseudomonas syringae).

B. A gene conferring resistance to a pest, such as soybean cystnematode. See e.g., PCT Application WO 96/30517; PCT Application WO93/19181.

C. A Bacillus thuringiensis protein, a derivative thereof or a syntheticpolypeptide modeled thereon. See, for example, Geiser et al., Gene48:109 (1986), who disclose the cloning and nucleotide sequence of a Btδ-endotoxin gene. Moreover, DNA molecules encoding δ-endotoxin genes canbe purchased from American Type Culture Collection, Manassas, Va., forexample, under ATCC Accession Nos. 40098, 67136, 31995 and 31998.

D. A lectin. See, for example, the disclosure by Van Damme et al., PlantMolec. Biol. 24:25 (1994), who disclose the nucleotide sequences ofseveral Clivia miniata mannose-binding lectin genes.

E. A vitamin-binding protein such as avidin. See PCT applicationUS93/06487. The application teaches the use of avidin and avidinhomologues as larvicides against insect pests.

F. An enzyme inhibitor, for example, a protease or proteinase inhibitoror an amylase inhibitor. See, for example, Abe et al., J. Biol. Chem.262:16793 (1987) (nucleotide sequence of rice cysteine proteinaseinhibitor); Huub et al., Plant Molec. Biol. 21:985 (1993) (nucleotidesequence of cDNA encoding tobacco proteinase inhibitor I); Sumitani etal., Biosci. Biotech. Biochem. 57:1243 (1993) (nucleotide sequence ofStreptomyces nitrosporeus α-amylase inhibitor); and U.S. Pat. No.5,494,813 (Hepher and Atkinson, issued Feb. 27, 1996).

G. An insect-specific hormone or pheromone such as an ecdysteroid orjuvenile hormone, a variant thereof, a mimetic based thereon, or anantagonist or agonist thereof. See, for example, the disclosure byHammock et al., Nature 344:458 (1990), of baculovirus expression ofcloned juvenile hormone esterase, an inactivator of juvenile hormone.

H. An insect-specific peptide or neuropeptide which, upon expression,disrupts the physiology of the affected pest. For example, see thedisclosures of Regan, J. Biol. Chem. 269:9 (1994) (expression cloningyields DNA coding for insect diuretic hormone receptor); and Pratt etal., Biochem. Biophys. Res. Comm. 163:1243 (1989) (an allostatin isidentified in Diploptera puntata). See also U.S. Pat. No. 5,266,317 toTomalski et al., who disclose genes encoding insect-specific, paralyticneurotoxins.

I. An insect-specific venom produced in nature by a snake, a wasp, etc.For example, see Pang et al., Gene 116:165 (1992), for disclosure ofheterologous expression in plants of a gene coding for a scorpioninsectotoxic peptide.

J. An enzyme responsible for a hyperaccumulation of a monoterpene, asesquiterpene, a steroid, hydroxamic acid, a phenylpropanoid derivativeor another non-protein molecule with insecticidal activity.

K. An enzyme involved in the modification, including thepost-translational modification, of a biologically active molecule; forexample, a glycolytic enzyme, a proteolytic enzyme, a lipolytic enzyme,a nuclease, a cyclase, a transaminase, an esterase, a hydrolase, aphosphatase, a kinase, a phosphorylase, a polymerase, an elastase, achitinase and a glucanase, whether natural or synthetic. See PCTapplication WO 93/02197 in the name of Scott et al., which discloses thenucleotide sequence of a callase gene. DNA molecules which containchitinase-encoding sequences can be obtained, for example, from the ATCCunder Accession Nos. 39637 and 67152. See also Kramer et al., InsectBiochem. Molec. Biol. 23:691 (1993), who teach the nucleotide sequenceof a cDNA encoding tobacco hornworm chitinase; and Kawalleck et al.,Plant Molec. Biol. 21:673 (1993), who provide the nucleotide sequence ofthe parsley ubi4-2 polyubiquitin gene.

L. A molecule that stimulates signal transduction. For example, see thedisclosure by Botella et al., Plant Molec. Biol. 24:757 (1994), ofnucleotide sequences for mung bean calmodulin cDNA clones; and Griess etal., Plant Physiol. 104:1467 (1994), who provide the nucleotide sequenceof a maize calmodulin cDNA clone.

M. A hydrophobic moment peptide. See PCT application WO 95/16776(disclosure of peptide derivatives of Tachyplesin which inhibit fungalplant pathogens) and PCT application WO 95/18855 (teaches syntheticantimicrobial peptides that confer disease resistance).

N. A membrane permease, a channel former or a channel blocker. Forexample, see the disclosure of Jaynes et al., Plant Sci. 89:43 (1993),of heterologous expression of a cecropin-β, lytic peptide analog torender transgenic tobacco plants resistant to Pseudomonas solanacearum.

O. A viral-invasive protein or a complex toxin derived therefrom. Forexample, the accumulation of viral coat proteins in transformed plantcells imparts resistance to viral infection and/or disease developmenteffected by the virus from which the coat protein gene is derived, aswell as by related viruses. See Beachy et al., Ann. rev. Phytopathol.28:451 (1990). Coat protein-mediated resistance has been conferred upontransformed plants against alfalfa mosaic virus, cucumber mosaic virus,tobacco streak virus, potato virus X, potato virus Y, tobacco etchvirus, tobacco rattle virus and tobacco mosaic virus. Id.

P. An insect-specific antibody or an immunotoxin derived therefrom.Thus, an antibody targeted to a critical metabolic function in theinsect gut would inactivate an affected enzyme, killing the insect. Cf.Taylor et al., Abstract #497, Seventh Int'l Symposium on MolecularPlant-Microbe Interactions (Edinburgh, Scotland) (1994) (enzymaticinactivation in transgenic tobacco via production of single-chainantibody fragments).

Q. A virus-specific antibody. See, for example, Tavladoraki et al.,Nature 366:469 (1993), who show that transgenic plants expressingrecombinant antibody genes are protected from virus attack.

R. A developmental-arrestive protein produced in nature by a pathogen ora parasite. Thus, fungal endo α-1,4-D-polygalacturonases facilitatefungal colonization and plant nutrient release by solubilizing plantcell wall homo-α-1,4-D-galacturonase. See Lamb et al., Bio/Technology10:1436 (1992). The cloning and characterization of a gene which encodesa bean endopolygalacturonase-inhibiting protein is described by Toubartet al., Plant J. 2:367 (1992).

S. A developmental-arrestive protein produced in nature by a plant. Forexample, Logemann et al., Bio/Technology 10:305 (1992), have shown thattransgenic plants expressing the barley ribosome-inactivating gene havean increased resistance to fungal disease.

2. Genes That Confer Resistance to an Herbicide:

A. An herbicide that inhibits the growing point or meristem, such as animidazolinone or a sulfonylurea. Exemplary genes in this category codefor mutant ALS and AHAS enzyme as described, for example, by Lee et al.,EMBO J. 7:1241 (1988); and Mild et al., Theor. Appl. Genet. 80:449(1990), respectively.

B. Glyphosate (resistance conferred by, e.g., mutant5-enolpyruvylshikimate-3-phosphate synthase (EPSPs) genes (via theintroduction of recombinant nucleic acids and/or various forms of invivo mutagenesis of native EPSPs genes), aroA genes and glyphosateacetyl transferase (GAT) genes, respectively), other phosphono compoundssuch as glufosinate (phosphinothricin acetyl transferase (PAT) genesfrom Streptomyces species, including Streptomyces hygroscopicus andStreptomyces viridichromogenes), and pyridinoxy or phenoxy proprionicacids and cyclohexones (ACCase inhibitor-encoding genes), See, forexample, U.S. Pat. No. 4,940,835 to Shah, et al. and U.S. Pat. No.6,248,876 to Barry et al., which disclose nucleotide sequences of formsof EPSPs which can confer glyphosate resistance to a plant. A DNAmolecule encoding a mutant aroA gene can be obtained under ATCCaccession number 39256, and the nucleotide sequence of the mutant geneis disclosed in U.S. Pat. No. 4,769,061 to Comai. European patentapplication No. 0 333 033 to Kumada et al., and U.S. Pat. No. 4,975,374to Goodman et al., disclose nucleotide sequences of glutamine synthetasegenes which confer resistance to herbicides such as L-phosphinothricin.The nucleotide sequence of a PAT gene is provided in Europeanapplication No. 0 242 246 to Leemans et al., DeGreef et al.,Bio/Technology 7:61 (1989), describe the production of transgenic plantsthat express chimeric bar genes coding for PAT activity. Exemplary ofgenes conferring resistance to phenoxy proprionic acids andcyclohexones, such as sethoxydim and haloxyfop are the Acc1-S1, Acc1-S2and Acc1-S3 genes described by Marshall et al., Theor. Appl. Genet.83:435 (1992). GAT genes capable of conferring glyphosate resistance aredescribed in WO 2005012515 to Castle et al. Genes conferring resistanceto 2,4-D, fop and pyridyloxy auxin herbicides are described in WO2005107437 and U.S. patent application Ser. No. 11/587,893, bothassigned to Dow AgroSciences LLC.

C. An herbicide that inhibits photosynthesis, such as a triazine (psbAand gs+ genes) or a benzonitrile (nitrilase gene). Przibila et al.,Plant Cell 3:169 (1991), describe the transformation of Chlamydomonaswith plasmids encoding mutant psbA genes. Nucleotide sequences fornitrilase genes are disclosed in U.S. Pat. No. 4,810,648 to Stalker, andDNA molecules containing these genes are available under ATCC AccessionNos. 53435, 67441, and 67442. Cloning and expression of DNA coding for aglutathione S-transferase is described by Hayes et al., Biochem. J.285:173 (1992).

3. Genes that Confer or Contribute to a Value-Added Trait, Such as:

A. Modified fatty acid metabolism, for example, by transforming a plantwith an antisense gene of stearyl-ACP desaturase to increase stearicacid content of the plant. See Knultzon et al., Proc. Natl. Acad. Sci.U.S.A. 89:2624 (1992).

B. Decreased phytate content—1) Introduction of a phytase-encoding genewould enhance breakdown of phytate, adding more free phosphate to thetransformed plant. For example, see Van Hartingsveldt et al., Gene127:87 (1993), for a disclosure of the nucleotide sequence of anAspergillus niger phytase gene. 2) A gene could be introduced thatreduced phytate content. In maize for example, this could beaccomplished by cloning and then reintroducing DNA associated with thesingle allele which is responsible for maize mutants characterized bylow levels of phytic acid. See Raboy et al., Maydica 35:383 (1990).

C. Modified carbohydrate composition effected, for example, bytransforming plants with a gene coding for an enzyme that alters thebranching pattern of starch. See Shiroza et al., J. Bacteriol. 170:810(1988) (nucleotide sequence of Streptococcus mutantsfructosyltransferase gene); Steinmetz et al., Mol. Gen. Genet. 20:220(1985) (nucleotide sequence of Bacillus subtilis levansucrase gene); Penet al., Bio/Technology 10:292 (1992) (production of transgenic plantsthat express Bacillus lichenifonnis α-amylase); Elliot et al., PlantMolec. Biol. 21:515 (1993) (nucleotide sequences of tomato invertasegenes); Sogaard et al., J. Biol. Chem. 268:22480 (1993) (site-directedmutagenesis of barley α-amylase gene); and Fisher et al., Plant Physiol.102:1045 (1993) (maize endosperm starch branching enzyme II).

Methods for Sunflower Transformation

Numerous methods for plant transformation have been developed includingbiological and physical plant transformation protocols. See, forexample, Mild et al., “Procedures for Introducing Foreign DNA intoPlants” in Methods in Plant Molecular Biology and Biotechnology, B. R.Glick and J. E. Thompson, Eds. (CRC Press, Inc., Boca Raton, 1993) pages67-88. In addition, expression vectors and in vitro culture methods forplant cell or tissue transformation and regeneration of plants areavailable. See, for example, Gruber et al., “Vectors for PlantTransformation” in Methods in Plant Molecular Biology and Biotechnology,B. R. Glick and J. E. Thompson, Eds. (CRC Press, Inc., Boca Raton, 1993)pages 89-119.

A. Agrobacterium-mediated Transformation—One method for introducing anexpression vector into plants is based on the natural transformationsystem of Agrobacterium. See, for example, Horsch et al., Science227:1229 (1985). A. tumefaciens and A. rhizogenes are plant pathogenicsoil bacteria which genetically transform plant cells. The Ti and Riplasmids of A. tumefaciens and A. rhizogenes, respectively, carry genesresponsible for genetic transformation of the plant. See, for example,C. I. Kado, Crit. Rev. Plant Sci. 10:1 (1991). Descriptions ofAgrobacterium vector systems and methods for Agrobacterium-mediated genetransfer are provided by Gruber et al., supra, Mild et al., supra, andMoloney et al., Plant Cell Reports 8:238 (1989). See also, U.S. Pat. No.5,563,055 (Townsend and Thomas), issued Oct. 8, 1996.

B. Direct Gene Transfer—Several methods of plant transformation,collectively referred to as direct gene transfer, have been developed asan alternative to Agrobacterium-mediated transformation. A generallyapplicable method of plant transformation is microprojectile-mediatedtransformation wherein DNA is carried on the surface of microprojectilesmeasuring 1 to 4 μm. The expression vector is introduced into planttissues with a biolistic device that accelerates the microprojectiles tospeeds of 300 to 600 m/s which is sufficient to penetrate plant cellwalls and membranes. Sanford et al., Part. Sci. Technol. 5:27 (1987); J.C. Sanford, Trends Biotech. 6:299 (1988); Klein et al., Bio/Technology6:559-563 (1988); J. C. Sanford, Physiol. Plant 7:206 (1990); Klein etal., Biotechnology 10:268 (1992). See also U.S. Pat. No. 5,015,580(Christou, et al.), issued May 14, 1991; U.S. Pat. No. 5,322,783 (Tomes,et al.), issued Jun. 21, 1994.

Another method for physical delivery of DNA to plants is sonication oftarget cells. Zhang et al., Bio/Technology 9:996 (1991). Alternatively,liposome and spheroplast fusion have been used to introduce expressionvectors into plants. Deshayes et al., EMBO. J., 4:2731 (1985); Christouet al., Proc. Natl. Acad. Sci. U.S.A. 84:3962 (1987). Direct uptake ofDNA into protoplasts using CaCl₂ precipitation, polyvinyl alcohol orpoly-L-ornithine has also been reported. Hain et al., Mol. Gen. Genet.199:161 (1985); and Draper et al., Plant Cell Physiol. 23:451 (1982).Electroporation of protoplasts and whole cells and tissues have alsobeen described. Donn et al., In Abstracts of VIIth InternationalCongress on Plant Cell and Tissue Culture IAPTC, A2-38, p 53 (1990);D'Halluin et al., Plant Cell 4:1495-1505 (1992); and Spencer et al.,Plant Mol. Biol. 24:51-61 (1994).

Following transformation of sunflower target tissues, expression of theabove-described selectable marker genes allows for preferentialselection of transformed cells, tissues and/or plants, usingregeneration and selection methods now well known in the art.

The foregoing methods for transformation would typically be used forproducing a transgenic variety. The transgenic variety could then becrossed, with another (non-transformed or transformed) variety, in orderto produce a new transgenic variety. Alternatively, a genetic traitwhich has been engineered into a particular sunflower cultivar using theforegoing transformation techniques could be moved into another cultivarusing traditional backcrossing techniques that are well known in theplant breeding arts. For example, a backcrossing approach could be usedto move an engineered trait from a public, non-elite variety into anelite variety, or from a variety containing a foreign gene in its genomeinto a variety or varieties which do not contain that gene. As usedherein, “crossing” can refer to a simple X by Y cross, or the process ofbackcrossing, depending on the context.

Tissue Culture of Sunflowers

Further production of the 503A/B cultivar can occur by self-pollinationor by tissue culture and regeneration. Tissue culture of various tissuesof sunflower and regeneration of plants therefrom is known. Furtherreproduction of the variety can occur by tissue culture andregeneration. Tissue culture of various tissues of soybeans andregeneration of plants therefrom is well known and widely published. Forexample, reference may be had to T. Komatsuda et al., “Genotype XSucrose Interactions for Somatic Embryogenesis in Soybeans,” Crop Sci.31:333-337 (1991); P. A. Stephens et al., “Agronomic Evaluation ofTissue-Culture-Derived Soybean Plants,” Theor. Appl. Genet. (1991)82:633-635; T. Komatsuda et al., “Maturation and Germination of SomaticEmbryos as Affected by Sucrose and Plant Growth Regulators in SoybeansGlycine gracilis Skvortz and Glycine max (L.) Merr.” Plant Cell, Tissueand Organ Culture, 28:103-113 (1992); S. Dhir et al., “Regeneration ofFertile Plants from Protoplasts of Soybean (Glycine max L. Merr.);Genotypic Differences in Culture Response,” Plant Cell Reports (1992)11:285-289; P. Pandey et al., “Plant Regeneration from Leaf andHypocotyl Explants of Glycine-wightii (W. and A.) VERDC. var.longicauda,” Japan J. Breed. 42:1-5 (1992); and K. Shetty et al.,“Stimulation of In Vitro Shoot Organogenesis in Glycine max (Merrill.)by Allantoin and Amides,” Plant Science 81:245-251 (1992). Thedisclosures of U.S. Pat. No. 5,024,944 issued Jun. 18, 1991 to Collinset al., and U.S. Pat. No. 5,008,200 issued Apr. 16, 1991 to Ranch etal., are hereby incorporated herein in their entirety by reference.Thus, another aspect of this invention is to provide cells which upongrowth and differentiation produce sunflower plants having thephysiological and morphological characteristics of sunflower variety503A/B.

As used herein, the term “tissue culture” indicates a compositioncomprising isolated cells of the same or a different type or acollection of such cells organized into parts of a plant. Exemplarytypes of tissue cultures are protoplasts, calli, plant clumps, and plantcells that can generate tissue culture that are intact in plants orparts of plants, such as embryos, pollen, flowers, seeds, pods, leaves,stems, roots, root tips, anthers, and the like. Means for preparing andmaintaining plant tissue culture are well known in the art. By way ofexample, a tissue culture comprising organs has been used to produceregenerated plants. U.S. Pat. Nos. 5,959,185, 5,973,234 and 5,977,445,describe certain techniques, the disclosures of which are incorporatedherein by reference.

Single-Gene Converted (Conversion) Plants

When the term “sunflower plant” is used in the context of the presentinvention, this also includes any single gene conversions of thatvariety. The term “single gene converted plant” as used herein refers tothose sunflower plants which are developed by a plant breeding techniquecalled backcrossing, or via genetic engineering, wherein essentially allof the desired morphological and physiological characteristics of avariety are recovered in addition to the single gene transferred intothe variety via the backcrossing technique. Backcrossing methods can beused with the present invention to improve or introduce a characteristicinto the variety. The term “backcrossing” as used herein refers to therepeated crossing of a hybrid progeny back to the recurrent parent,i.e., backcrossing 1, 2, 3, 4, 5, 6, 7, 8 or more times to the recurrentparent. The parental sunflower plant which contributes the gene for thedesired characteristic is termed the “nonrecurrent” or “donor parent.”This terminology refers to the fact that the nonrecurrent parent is usedone time in the backcross protocol and therefore does not recur. Theparental sunflower plant to which the gene or genes from thenonrecurrent parent are transferred is known as the recurrent parent asit is used for several rounds in the backcrossing protocol (Poehlman &Sleper, 1994; Fehr, 1987). In a typical backcross protocol, the originalvariety of interest (recurrent parent) is crossed to a second variety(nonrecurrent parent) that carries the single gene of interest to betransferred. The resulting progeny from this cross are then crossedagain to the recurrent parent and the process is repeated until asunflower plant is obtained wherein essentially all of the desiredmorphological and physiological characteristics of the recurrent parentare recovered in the converted plant, in addition to the singletransferred gene from the nonrecurrent parent.

The selection of a suitable recurrent parent is an important step for asuccessful backcrossing procedure. The goal of a backcross protocol isto alter or substitute a single trait or characteristic in the originalvariety. To accomplish this, a single gene of the recurrent variety ismodified or substituted with the desired gene from the nonrecurrentparent, while retaining essentially all of the rest of the desiredgenetic, and therefore the desired physiological and morphological,constitution of the original variety. The choice of the particularnonrecurrent parent will depend on the purpose of the backcross. One ofthe major purposes is to add some commercially desirable, agronomicallyimportant trait to the plant. The exact backcrossing protocol willdepend on the characteristic or trait being altered to determine anappropriate testing protocol. Although backcrossing methods aresimplified when the characteristic being transferred is a dominantallele, a recessive allele may also be transferred. In this instance itmay be necessary to introduce a test of the progeny to determine if thedesired characteristic has been successfully transferred.

Many single gene traits have been identified that are not regularlyselected for in the development of a new variety but that can beimproved by backcrossing techniques. Single gene traits may or may notbe transgenic, examples of these traits include but are not limited to,male sterility, waxy starch, herbicide resistance, resistance forbacterial, fungal, or viral disease, insect resistance, male fertility,enhanced nutritional quality, industrial usage, yield stability andyield enhancement. These genes are generally inherited through thenucleus. Several of these single gene traits are described in U.S. Pat.Nos. 5,959,185, 5,973,234 and 5,977,445, the disclosures of which arespecifically hereby incorporated by reference.

This invention also is directed to methods for producing a sunflowerplant by crossing a first parent sunflower plant with a second parentsunflower plant wherein the first or second parent sunflower plant is asunflower plant of the variety 503A/B. Further, both first and secondparent sunflower plants can come from the sunflower variety 503A/B.Thus, any such methods using the sunflower variety 503A/B are part ofthis invention: selfing, backcrosses, hybrid production, crosses topopulations, and the like. All plants produced using sunflower variety503A/B as a parent are within the scope of this invention, includingthose developed from varieties derived from sunflower variety 503A/B.Advantageously, the sunflower variety could be used in crosses withother, different, sunflower plants to produce first generation (F₁)sunflower hybrid seeds and plants with superior characteristics. Thevariety of the invention can also be used for transformation whereexogenous genes are introduced and expressed by the variety of theinvention. Genetic variants created either through traditional breedingmethods using variety 503A/B or through transformation of 503A/B by anyof a number of protocols known to those of skill in the art are intendedto be within the scope of this invention.

The invention is also directed to Sunflower meal from seeds of an elitesunflower variety.

Oxidative Stability

Stability can be defined as the resistance of a vegetable oil tooxidation and to the resulting deterioration due to the generation ofproducts causing rancidity and decreasing food quality. Tests foroxidative stability attempt to accelerate the normal oxidation processto yield results that can be translated into quality parameters fordifferent food ails and to predict their shelf lives. Stability methodsare also useful to evaluate antioxidants and their effects on protectionof foods against lipid oxidation.

Lipid oxidation in food products develops slowly initially, and thenaccelerates at later stages during storage. The induction period isdefined as the time to reach a constant percent oxidation of the fat asrelated to the end of shelf life. The induction period is measuredeither as the time required for a sudden change in rate of oxidation, orby estimating the intersection point between the initial and final ratesof oxidation. For vegetable oils containing linoleic and linolenic acid,such as soybean and sunflower oils, the end-points for acceptabilitywill occur at relatively low levels of oxidation (peroxide valuesbetween 1 and 10 Meq/kg).

Factors Affecting Oxidative Stability

The difference in stability between different vegetable oils is due totheir different fatty acid profiles, the effect of processing, initiallevels of oxidation at the start of the storage period, and otherfactors including, minor components, including the presence of metalimpurities, formulation, packaging and environmental storage conditions.From the crude stage to different stages of processing of vegetableoils, some oxidation can take place that will affect the subsequentoxidative stability of the final oil product during storage.

Oxidative Stability Methods

To estimate the oxidative stability of a fat to oxidation, the sample issubjected to an accelerated oxidation test under standardized conditionsand a suitable end-point is chosen to determine the level of oxidativedeterioration. Methods involving elevated temperatures include:

1. Schaal Oven Test

The sample is heated at 50 to 60° C. until it reaches a suitableend-point based on peroxide value or carbonyl value such as theanisidine value. The results of this test correlate best with actualshelf life because the peroxide value end-point of 10 represents arelatively low degree of oxidation. See, limiting peroxide value insection D below.

2. Active Oxygen Method (AOM), Rancimat and Oxidation Stability Index(OSI). See, e.g., U.S. Pat. No. 5,339,294 to Matlock et al., AOCS Method12b-92; and M. W. Laubli and P. A. Bruttel, JOACS 63:792-795 (1986).

Air is bubbled through a sample of oil in special test tubes heated at98-100° C. and the progress of oxidation is followed by peroxide valuedetermination in the AOM test, and by conductivity measurements in theRancimat and OSI tests. The automated Rancimat and OSI tests may be runat temperatures ranging from 100-140° C., and the effluent gases are ledthrough a vessel containing deionized water and the increase inconductivity measured are due to the formation of volatile organic acids(mainly formic acid) by thermal oxidation. The OSI is defined as thetime point in hours of maximum change of the rate of oxidation based onconductivity.

D. Methods to Determine Oxidation—The peroxide value of oils is ameasure of oxidation that is useful for samples that are oxidized torelatively low levels (peroxide values of less than 50), and underconditions sufficiently mild so that the hydroperoxides, which are theprimary products formed by oxidation, are not markedly decomposed. Alimiting peroxide value of 10 meq/kg was specified for refined oils byFAQ/WHO standards (Joint FAQ/WHO Food Standard Program CodexAlimentarius Commission, Report of 16th session of Committee on Fats andOils, London, 1999).

The anisidine test measures high molecular weight saturated andunsaturated carbonyl compounds in oils. The test provides usefulinformation on non-volatile carbonyl compounds formed in oils duringprocessing of oils containing linolenate. The Totox value (anisidinevalue+2 times peroxide value) is used as an empirical measure of theprecursor non-volatile carbonyl compounds present in processed oils plusany further oxidation products developed after storage.

Deposit Information

A deposit of the Dow AgroSciences proprietary sunflower cultivar 503A/Bdisclosed above and recited in the appended claims has been made withthe American Type Culture Collection (ATCC), 10801 University Boulevard,Manassas, Va. 20110. The date of deposit was May 29, 2009. The depositof 2500 seeds were taken from the same deposit maintained by DowAgroSciences LLC since prior to the filing date of this application. Allrestrictions upon the deposit have been removed, and the deposit isintended to meet all of the requirements of 37 C.F.R. Sections1.801-1.809. The ATCC accession number is PTA-10098. The deposit will bemaintained in the depository for a period of 30 years, or 5 years afterthe last request, or for the effective life of the patent, whichever islonger, and will be replaced as necessary during that period.

While a number of exemplary aspects and embodiments have been discussedabove, those of skill in the art will recognize certain modifications,permutations, additions and sub-combinations thereof. It is thereforeintended that the following appended claims and claims hereafterintroduced are interpreted to include all such modifications,permutations, additions and sub-combinations as are within their truespirit and scope.

All references, including publications, patents, and patentapplications, cited herein are hereby incorporated by reference to thesame extent as if each reference were individually and specificallyindicated to be incorporated by reference and were set forth in itsentirety herein. The references discussed herein are provided solely fortheir disclosure prior to the filing date of the present application.Nothing herein is to be construed as an admission that the inventors arenot entitled to antedate such disclosure by virtue of prior invention.

1. A seed of sunflower inbred line designated 503A/B, representativeseed of said line having been deposited under ATCC Accession No.PTA-10098.
 2. A sunflower plant, or part thereof, produced by growingthe seed of claim
 1. 3. Pollen of the plant of claim
 2. 4. An ovule ofthe plant of claim
 2. 5. A sunflower plant, or parts thereof, having allof the physiological and morphological characteristics of the sunflowerplant of claim
 2. 6. A tissue culture of regenerable cells from thesunflower plant of claim
 2. 7. The tissue culture according to claim 6,wherein a cell or protoplast of the tissue culture is derived from aplant part selected from the group consisting of leaves, pollen,embryos, roots, root tips, anthers, flowers, and stalks.
 8. A sunflowerplant regenerated from the tissue culture of claim 6, wherein theregenerated plant has all the morphological and physiologicalcharacteristics of inbred line 503A/B, representative seed of said line503A/B having been deposited under ATCC Accession No. PTA-10098.
 9. Asunflower plant with all of the physiological and morphologicalcharacteristics of inbred line 503A/B, wherein said sunflower plant isproduced by a tissue culture process using the sunflower plant of claim5 as the starting material for said process.
 10. A method for producinga hybrid sunflower seed, wherein said method comprises crossing a firstinbred parent sunflower plant with a second inbred parent sunflowerplant and harvesting the resultant hybrid sunflower seed, wherein saidfirst inbred parent sunflower plant or said second inbred parentsunflower plant is the sunflower plant of claim
 2. 11. The methodaccording to claim 10, wherein first inbred parent sunflower is 500B(1);and wherein the second inbred parent sunflower is 68204B.
 12. The methodaccording to claim 10, wherein first inbred parent sunflower is 503A/B;and wherein the second inbred parent sunflower is selected from thegroup consisting of 319R, 318R, and 222R.
 13. The hybrid sunflower seedss671, s674, and s878 are produced by the method of claim
 12. 14. Amethod for producing a male-sterile sunflower plant comprisingtransforming the sunflower plant of claim 2 with a nucleic acid moleculethat confers male sterility.
 15. A male sterile sunflower plant producedby the method of claim
 14. 16. A method of producing an herbicideresistant sunflower plant comprising transforming the sunflower plant ofclaim 2 with a transgene that confers herbicide resistance.
 17. Anherbicide resistant sunflower plant produced by the method of claim 16.18. The sunflower plant of claim 17, wherein the transgene confersresistance to an herbicide selected from the group consisting of,imidazolinone, sulfonylurea, glyphosate, glufosinate,L-phosphinothricin, triazine and benzonitrile.
 19. The sunflower plantof claim 17, wherein the transgene confers resistance to the herbicideimidazolinone.
 20. A method of producing a disease resistant sunflowerplant comprising transforming the sunflower plant of claim 2 with atransgene that confers disease resistance.
 21. A disease resistantsunflower plant produced by the method of claim
 20. 22. A method ofintroducing a desired trait into sunflower inbred line 503A/B, whereinthe method comprises: (a) crossing 503A/B plants grown from 503A/B seedrepresentative seed of which has been deposited under ATCC Accession No.PTA-10098, with plants of another sunflower line that comprise a desiredtrait to produce progeny plants, wherein the desired trait is selectedfrom the group consisting of male sterility, herbicide resistance,insect resistance, disease resistance and oil content; (b) selectingprogeny plants that have the desired trait to produce selected progenyplants; (c) crossing the selected progeny plants with the 503A/B plantsto produce backcross progeny plants; (d) selecting for backcross progenyplants that have the desired trait and physiological and morphologicalcharacteristics of sunflower inbred line 503A/B to produce selectedbackcross progeny plants; and (e) repeating steps (c) and (d) three moretimes in succession to produce selected fourth or higher backcrossprogeny plants that comprise the desired trait and all of thephysiological and morphological characteristics of sunflower inbred line503A/B.
 23. A plant produced by the method of claim 22, wherein theplant has the desired trait and all of the physiological andmorphological characteristics of sunflower inbred line 503A/B.
 24. Theplant of claim 23, wherein the desired trait is male sterility and thetrait conferred by the cytoplasmic nucleic acid molecule that confersmale sterility.
 25. The plant of claim 23, wherein the desired trait isherbicide resistance to an herbicide selected from the group consistingof, imidazolinone, sulfonylurea, glyphosate, glufosinate,L-phosphinothricin, triazine and benzonitrile.
 26. The plant of claim23, wherein the desired trait is herbicide resistance to imidazolinone.